CHEMCELL · Chemical Biology in Reactors and Cells

Final Activity Report Summary - CHEMCELL (Chemical Biology in Reactors and Cells)

The CHEMCELL programme was established to train young scientists to tackle problems at the chemistry biology interface that would benefit both industry and wider society as a whole. In particular, two major challenges in a post-genomics era were identified as being: 1. To meet the increasing demand for novel, relevant, biological active compounds that can be used as drugs; 2. To deliver sensitive and quantitative extraction and detection methodologies for the high-throughput analysis of proteins and small molecules in research programmes aimed at determining the function of genes and the composition of the food we eat. Enzymes offer a number of major advantages over chemical catalysts, in terms of their efficiency and ability to perform exquisite chemistries and are becoming increasingly important as biocatalysts for the synthesis active pharmaceutical ingredients. By bringing together the disciplines of chemistry and biology in the CHEMCELL programme we have been able to make good progress towards improving the value of enzyme-catalysed reactions. By using porous inorganic solids with the right bulk and surface structures, we have successfully trapped enzymes in a way that makes them easy to use, effective as catalysts and stable on storage. The use of enzymes in industry is often impeded by their requirement for water as a solvent and to overcome this problem we have examined enzyme catalysis in protic ionic liquids (PILs), ammonium salts that are liquid at room temperature. CHEMCELL has shown that tailoring the structure of PILs can facilitate the performance of enzyme catalysis in these media, in the presence of low levels of water. The use of PILs as solvents for biocatalysis is attractive as they have low vapour pressure, are recyclable, and environmentally friendly. Plant enzymes capable of adding sugars to small molecules were investigated as biocatalysts for the preparation of a diverse range of natural products. This aspect of the CHEMCELL programme has opened up the possibility of designing novel routes for those applications that increasingly require green chemistry, such as those in the fine chemical, flavour, fragrance and food additive sectors. CHEMCELL also focused on the development of methods for fast characterisation and comparison of enzymes. Expenditure of enzymes and substrates was minimised by using capillaries as reaction and separation vessels, allowing the reactions to be carried in nanolitre volumes. The results have shown that a variety of enzyme assays can be set up by multiplexing capillaries and that area imaging UV detection provides unprecedented flexibility for applications in biochemical analysis. The development of technologies for the identification of health promoting components in plants was a key component of the CHEMCELL programme. CHEMCELL has developed a fast and reliable method for the analysis of fatty acids and individual triacylglycerols (TAGs) in plants. This was achieved through the synthesis of a novel standard reference set of TAGs and the development of a liquid chromatography-mass spectrometry analytical platform. This new methodology is giving insight into the regulation of oil synthesis and will provide valuable information for research programmes aimed at metabolic engineering of novel plant oils into oilseed crops. Carbohydrates are key compounds in plant metabolism; however, they are a particularly challenging class of compounds to identify and analyse. CHEMCELL research resulted in the development and optimisation of robust separations methods in combination with powerful mass spectrometric analysis for the simultaneous analysis of a range of carbohydrate metabolites. The use of mass spectrometry has also been important in CHEMCELL for the analysis important metabolites produced in plant glands called trichomes. We have investigated the effect of plant hormones on trichome development and on a trichome specific metabolic pathway producing acylsugars.

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